The Induced Membrane Technique—The Filling Matters: Evaluation of Different Forms of Membrane Filling with and without Bone Marrow Mononuclear Cells (BMC) in Large Femoral Bone Defects in Rats

Abstract

The Masquelet technique is used to treat large bone defects; it is a two-stage procedure based on an induced membrane. To improve the induced membrane process, demineralized bone matrix in granular (GDBM) and fibrous form (f-DBM) was tested with and without bone marrow mononuclear cells (BMC) as filling of the membrane against the gold standard filling with syngeneic cancellous bone (SCB). A total of 65 male Sprague–Dawley rats obtained a 5 mm femoral defect. These defects were treated with the induced membrane technique and filled with SCB, GDBM, or f-DBM, with or without BMC. After a healing period of eight weeks, the femurs were harvested and submitted for histological, radiological, and biomechanical analyses. The fracture load in the defect zone was lower compared to SCB in all groups. However, histological analysis showed comparable new bone formation, bone mineral density, and cartilage proportions and vascularization. The results suggest that f-DBM in combination with BMC and the induced membrane technique cannot reproduce the very good results of this material in large, non-membrane coated bone defects, nevertheless it supports the maturation of new bone tissue locally. It can be concluded that BMC should be applied in lower doses and inflammatory cells should be removed from the cell preparation before implantation.

Systematic literature review of in vivo rat femoral defect models using biomaterials to improve the induced membrane technique: a comprehensive analysis

“…The mean score on the ARRIVE Essential 10 checklist was 7.03 ± 0.97 (minimum–maximum 5–8) and the mean total score was 14.6 ± 1.8 (minimum–maximum 11–18) (Supplementary Table 1). The frequently missing items of the Essential Set were the description of the randomization method that was done partially or totally in 13 (65%) studies ( 8 , 9 , 10 , 11 , 12 , 13 , 14 , 15 , 16 , 17 , 18 , 19 , 20 ) and the blinding assessment of the study’s endpoints that were only done in one (5%) study ( 12 ). Finally, the a priori calculation of the number of animals needed for statistical comparison was only done in 5 (25%) studies ( 10 , 12 , 17 , 20 , 21 ).…”

“…The most frequent strain was the outbred Sprague-Dawley rat (16/20) ( 8 , 10 , 11 , 12 , 13 , 14 , 15 , 16 , 17 , 19 , 21 , 22 , 23 , 24 , 25 , 26 ), followed by Wistar (3/20) ( 9 , 20 , 27 ) and one study used an inbred 344 Fischer strain ( 18 ). Only 2/20 studies used female rats (Wistar) ( 9 , 27 ).…”

“…The mean length of the femoral defect was 7 ± 2.2 mm (minimum–maximum 4–10). Stabilization of femoral defect was principally achieved with one plate and 4 screws (13/20) ( 9 , 10 , 12 , 13 , 14 , 17 , 18 , 19 , 20 , 22 , 23 , 24 , 27 ) or 6 screws (1/20) ( 26 ), followed by external fixation (4/20) ( 11 , 15 , 21 , 25 ) and intramedullary nailing (2/20) ( 8 , 16 ). Two studies used an additional cerclage wiring ( 9 , 26 ).…”

“…Two studies used an additional cerclage wiring ( 9 , 26 ). The defect was grafted in 14/20 studies and consisted of either bone allograft ( 11 , 13 , 15 , 17 , 18 , 19 , 24 ), autograft ( 14 , 25 ) or calcium phosphate material ( 8 , 12 , 16 , 17 , 19 , 23 , 24 , 26 ) that could be compared. In this model, the time of IM formation was most frequently found at 4 weeks (14/20) (extremes 2–11 weeks), but only three studies analyzed the formation of IM kinetically at several time points (from 2 to 8 weeks) ( 10 , 15 , 22 ) (Supplementary Table 2).…”

“…The main strategy consisted of an optimization of the conventional two-stage IMT (13/20) ( 8 , 10 , 11 , 14 , 15 , 17 , 18 , 19 , 20 , 21 , 24 , 25 , 27 ). A part of them (4/13) loaded the PMMA spacer with bioactive molecules such as antibiotics and growth factors ( 10 , 18 , 20 , 25 ).…”